title: Time-resolved fluorescence spectroscopy investigation of the effect of 4-hydroxynonenal on endogenous NAD(P)H in living cardiac myocytes.
abstract: Lipid peroxidation is a major biochemical consequence of the oxidative deterioration of polyunsaturated lipids in cell membranes and causes damage to membrane integrity and loss of protein function. 4-hydroxy-2-nonenal (HNE), one of the most reactive products of n-6 polyunsaturated fatty acid peroxidation of membrane phospholipids, has been shown to be capable of affecting both nicotinamide adenine dinucleotide (phosphate) reduced [NAD(P)H] as well as NADH production. However, the understanding of its effects in living cardiac cells is still lacking. Our goal was to therefore investigate HNE effects on NAD(P)H noninvasively in living cardiomyocytes. Spectrally resolved lifetime detection of endogenous fluorescence, an innovative noninvasive technique, was employed. Individual fluorescence components were resolved by spectral linear unmixing approach. Gathered results revealed that HNE reduced the amplitude of both resolved NAD(P)H components in a concentration-dependent manner. In addition, HNE increased flavoprotein fluorescence and responsiveness of the NAD(P)H component ratio to glutathione reductase (GR) inhibitor. HNE also increased the percentage of oxidized nucleotides and decreased maximal NADH production. Presented data indicate that HNE provoked an important cell oxidation by acting on NAD(P)H regulating systems in cardiomyocytes. Understanding the precise role of oxidative processes and their products in living cells is crucial for finding new noninvasive tools for biomedical diagnostics of pathophysiological states.
authors.affiliation: International Laser Center, Department of Biophotonics, Bratislava, Slovakia. alzbeta.chorvatova@ilc.sk
authors.forename: Alzbeta
authors.initials: A
authors.lastname: Chorvatova
authors.forename: Swida
authors.initials: S
authors.lastname: Aneba
authors.forename: Anton
authors.initials: A
authors.lastname: Mateasik
authors.forename: Dusan
authors.initials: D
authors.lastname: Chorvat
authors.forename: Blandine
authors.initials: B
authors.lastname: Comte
mesh_terms: D000447:Aldehydes / Q000494:pharmacology*; D000818:Animals; D004138:Dinitrocresols / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D005260:Female; D005420:Flavoproteins / Q000378:metabolism; D005980:Glutathione Reductase / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D015227:Lipid Peroxidation; D032383:Myocytes, Cardiac / Q000166:cytology / Q000378:metabolism*; D009249:NADP / Q000378:metabolism*; D010100:Oxygen / Q000378:metabolism; D010743:Phospholipids / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D017382:Reactive Oxygen Species; D013050:Spectrometry, Fluorescence
chemical_list: D000447:Aldehydes; D004138:Dinitrocresols; D005420:Flavoproteins; D010743:Phospholipids; D017382:Reactive Oxygen Species; C024132:4,6-dinitro-o-cresol; D009249:NADP; D005980:Glutathione Reductase; C027576:4-hydroxy-2-nonenal; D010100:Oxygen